In-Fusion™克隆技术在大肠杆菌表达载体平行构建中的应用。

Application of In-Fusion™ cloning for the parallel construction of E. coli expression vectors.

作者信息

Bird Louise E, Rada Heather, Flanagan John, Diprose Jonathan M, Gilbert Robert J C, Owens Raymond J

机构信息

Oxford Protein Production Facility-UK, Oxfordshire, UK.

出版信息

Methods Mol Biol. 2014;1116:209-34. doi: 10.1007/978-1-62703-764-8_15.

DOI:10.1007/978-1-62703-764-8_15

PMID:24395367

Abstract

In-Fusion™ cloning is a flexible DNA ligase-independent cloning technology that has wide-ranging uses in molecular biology. In this chapter we describe the protocols used in the OPPF-UK to design and construct expression vectors using In-Fusion™. Our method for small scale expression screening in Escherichia coli of constructs generated by In-Fusion™ is also outlined.

摘要

In-Fusion™克隆是一种灵活的不依赖DNA连接酶的克隆技术，在分子生物学中有广泛应用。在本章中，我们描述了英国植物表型分析设施（OPPF-UK）中使用In-Fusion™设计和构建表达载体的方案。我们还概述了对In-Fusion™产生的构建体在大肠杆菌中进行小规模表达筛选的方法。